Method for manufacturing a biological fluid sensor

ABSTRACT

The present invention presents a method of fabrication for a physiological sensor with electronic, electrochemical, and chemical components. The fabrication method comprises steps for manufacturing an apparatus comprising at least one electrochemical sensor, a microcontroller, and a transceiver. The fabrication process includes the steps of substrate fabrication, circuit fabrication, pick and place, reflow soldering, electrode fabrication, membrane fabrication, sealing and curing, layer bonding, and dressing. The physiological sensor is operable to analyze biological fluids such as sweat.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.16/791,430, filed Feb. 14, 2020, which is a continuation of U.S.application Ser. No. 15/487,046, filed Apr. 13, 2017, which is acontinuation-in-part of U.S. application Ser. No. 15/177,703, filed Jun.9, 2016, which is a continuation-in-part of U.S. application Ser. No.15/019,006, filed Feb. 9, 2016, which claims priority from U.S.Provisional Patent Application No. 62/130,047, filed Mar. 9, 2015, eachof which is incorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION 1. Field of the Invention

The present invention is generally directed to a method formanufacturing a device operable for collecting and analyzing biologicalfluid inputs, calculating biological fluid concentrations and ratios,and transmitting biological fluid values to a transceiver device.

2. Description of the Prior Art

Generally, biomarkers from biological fluid have significant prognosticand/or diagnostic utility, such as predicting disease, nutritionalimbalance, or psychological or physical stress; however, many of themost utilized biomarkers are collected from blood. The ability topredict events through non-invasive means, such as sweat detection,provides great utility to persons under physical stress, particularlyindividuals in the process of physical activity or exercise. The abilityto monitor sweat biomarkers in real time and continuously duringactivity allows an individual to make informed decisions regardinghydration, nutrition, and exertional status, and recovery, all variablesthat moderate physical performance.

For example, hydration status is a predictor of physical performance;dehydration as low as 1% of body mass can impair performance. Prior artdetection and treatment, as shown in FIG. 1 , is currently at the stagesof when symptoms present, performance degrades, and/or injury presents.Determining hydration through sweat biomarkers before dehydrationsymptoms present has many benefits, such as reducing fatigue, cramps,and headaches. Therefore, developing a device and system fornon-invasively obtaining biomarkers, such as through sweat, is needed.

Sweat contains a multitude of biomarkers; any substance aqueouslydissolvable in the blood can present in the sweat by way of eccrineglands. The sweat biomarkers can be small molecules (molecular weight<900 Daltons), proteins, metabolites, and/or electrolytes. Well-knownelectrolytes in sweat are sodium and potassium. As shown in FIG. 2 ,potassium concentration is not dependent upon sweat rate due to thepassive diffusive transport of potassium, while sodium and chlorideconcentrations in sweat are dependent upon sweat rate due to the activetransport of sodium. Thus, monitoring sodium or chloride concentrationsis an accurate, indirect means of indicating hydration status of anindividual. Therefore, developing a sweat biomarker device that cancommunicate to an individual real-time biomarker data is needed.

Prior art documents include the following:

U.S. Pat. No. 6,198,953 for method and system for continuous sweatcollection and analysis by inventors Webster, et al., filed Mar. 11,1999 and issued Mar. 6, 2001, is directed to a method and systemproviding especially for continuously obtaining and analyzing, on a realtime basis, sweat from a selected area of skin on the body of a person,especially a neonate, being diagnosed for cystic fibrosis, by causingsweating of the selected area of skin, by placing an electricallypositive iontophoretic electrode device of a set of said devices overthe selected area of skin preferably within a previously placedreceiving and holding device which, following the induction of sweat andremoval of the electrically positive iontophoretic electrode device,receives a sweat-sensing electrode device that continuously sendselectrical signals to sweat analysis circuitry for providing a digitalreadout of the ionic composition of the sweat.

U.S. Pat. No. 8,388,534 for apparatus for providing skin careinformation by measuring skin moisture content and method and medium forthe same by inventors Jang, et al., filed Sep. 24, 2007 and issued Mar.5, 2013, is directed to an apparatus for providing skin careinformation, the apparatus including: an electrode unit supplying avoltage to a user's skin and detecting a current signal in the user'sskin; a measurement control unit measuring the user's skin moisturecontent and sweat gland activity by using the detected current signal; adata calculation unit deriving skin moisture content information byusing the skin moisture content and the sweat gland activity, andgenerating skin care information corresponding to the skin moisturecontent information; and an information provider providing the user withthe generated skin care information.

U.S. Pat. No. 7,575,549 for an apparatus and method for increasing,monitoring, measuring, and controlling perspiratory water and solid lossat reduced ambient pressure by inventor Miller, filed Jul. 30, 2004 andissued Aug. 18, 2009, is directed to a device for increasing,monitoring, and measuring perspiration water and solid loss at reducedambient pressure, comprising a sealed chamber capable of maintainingless than atmospheric pressure for an extended period of time and agasket-sealed door accessing the chamber. An algorithm allowing forcontinuous calculations of sweat loss and fluid replacement requirementsof the occupant of the chamber is disclosed.

US Publication No. 20140330096 for method for performing a physiologicalanalysis with increased reliability by inventor Brunswick, filed Nov.12, 2012 and published Nov. 6, 2014, is directed to a method forperforming an electrophysiological analysis implemented in a system thatincludes: a series of electrodes to be placed on different regions ofthe human body; a DC voltage source controlled so as to produce DCvoltage pulses; a switching circuit for selectively connecting theactive electrodes to the voltage source, the active electrodes formingan anode and a cathode, and for connecting at least one otherhigh-impedance passive electrode used to measure the potential reachedby the body; and a measuring circuit for reading data representative ofthe current in the active electrodes, and data representative of thepotentials generated on at least certain high-impedance electrodes inresponse to the application of the pulses, the data allowing a value tobe determined for the electrochemical conductance of the skin. Themethod also regenerates a high-impedance electrode connected to thevoltage source as a cathode.

US Publication No. 20140350432 for assessment of relative proportions ofadrenergic and cholinergic nervous receptors with non-invasive tests byinventors Khalfallah, et al., filed Aug. 8, 2014 and published Nov. 27,2014, is directed to a system and method for assessing relativeproportions of cholinergic and adrenergic nervous receptors in apatient. The system includes: an anode, a cathode, and passive electrodefor placement on different regions of the patient body. The methodgenerally includes: applying DC voltage pulses of varying voltage valuesto stress sweat glands of the patient, collecting data representing thecurrent between the anode and the cathode and the potential of theanode, the cathode, and the passive electrode for each of the differentDC voltage, and computing data representing the electrochemical skinconductance of the patient. The computed data representing theelectromechanical skin conductance of the patient is reconciled withreference data from control patients having known relative proportionsof cholinergic and adrenergic nervous receptors. Thus, the relativeproportions of cholinergic and adrenergic nervous receptors in thepatient can be determined.

US Publication No. 20150019135 for motion sensor and analysis byinventors Kacyvenski, et al., filed Jun. 3, 2014 and published Jan. 15,2015, is directed to the performance of an individual being monitoredbased on measurements of a conformal sensor device. An example systemincludes a communication module to receive data indicative of ameasurement of at least one sensor component of the conformal sensordevice. The sensor component obtains measurement of acceleration datarepresentative of an acceleration proximate to the portion of theindividual. A comparison of a parameter computed based on the sensorcomponent measurement to a preset performance threshold value providesan indication of the performance of the individual.

The article Implementation of a Microfluidic Conductivity Sensor—APotential Sweat Electrolyte Sensing System for Dehydration Detection, byLiu, et al. in Conf Proc IEEE Eng Med Biol Soc, 2014:1678-81, discussesthe implementation of a microfluidic conductivity sensor—a potentialsweat electrolyte sensing system for dehydration detection.

US Publication No. 20130245388 for electronics for detection of acondition of tissue by inventors Rafferty, et al., filed Sep. 4, 2012and published Sep. 19, 2013, is directed to an apparatus for monitoringa condition of a tissue based on a measurement of an electrical propertyof the tissue. In an example, the electrical property of the tissue isperformed using an apparatus disposed above the tissue, where theapparatus includes at least two conductive structures, each having anon-linear configuration, where the at least two conductive structuresare disposed substantially parallel to each other. In another example,the electrical property of the tissue is performed using an apparatusdisposed above the tissue, where the apparatus includes at least oneinductor structure.

U.S. Pat. No. 5,690,893 for analyzer having sensor with memory device byinventors Ozawa, et al., filed Jun. 5, 1995 and issued Nov. 25, 1997, isdirected to an analyzer including an exchangeable and consumable elementsuch as a sensor, column or reagent the characteristic of whichspecifies an analyzing condition. The element is provided with anon-volatile semiconductor memory which holds an analyzing conditionadapted for the element as data. When the element is mounted on ananalyzer body, a controller of the analyzer reads the analyzingcondition from the memory to update an analyzing condition inherentlyprovided in the analyzer body. The result of analysis and/or operationalhistory information of use of the element may be written into the memorywith which the element is provided.

U.S. Pat. No. 8,241,697 for formation of immobilized biological layersfor sensing by inventors Collier, et al., filed Dec. 20, 2007 and issuedAug. 14, 2012, is directed to enzyme immobilization compositionscomprising: one or more enzymes, a humectant, an acrylic-based monomer,a water-soluble organic photo-initiator and a water-solubleacrylic-based cross-linker in a substantially homogeneous aqueousmixture. The publication also discloses to methods for forming sensorscomprising such compositions and to apparati for forming arrays ofimmobilized layers on an array of sensors by dispensing suchcompositions onto a substrate.

WIPO Publication No. WO2016061362 for sweat sensing device communicationsecurity and compliance by inventors Heikenfeld, et al., filed Oct. 15,2015 and published Jun. 16, 2016, is directed to addressing confoundingdifficulties involving continuous sweat analyte measurement.Specifically, the publication discloses: at least one component capableof monitoring whether a sweat sensing device is in sufficient contactwith a wearer's skin to allow proper device operation; at least onecomponent capable of monitoring whether the device is operating on awearer's skin; at least one means of determining whether the devicewearer is a target individual within a probability range; at least onecomponent capable of generating and communicating alert messages to thedevice user(s) related to: wearer safety, wearer physiologicalcondition, compliance with a requirement to wear a device, deviceoperation; compliance with a behavior requirement, or other purposesthat may be derived from sweat sensor data; and the ability to utilizeaggregated sweat sensor data that may be correlated with informationexternal to the device to enhance the predictive capabilities of thedevice.

Published article by Rose, et al. in IEEE Transactions on BiomedicalEngineering, Nov. 6, 2014, pages 1-9, discusses an adhesive RFID sensorpatch for monitoring of sweat electrolytes.

Although biomarkers in sweat are appreciated, specifically electrolytesand glucose, a system and method is still lacking that continuouslyanalyzes sweat biomarkers in real time and transmits data to a user,which informs the user of his or her health status.

SUMMARY OF THE INVENTION

The present invention provides a method of fabrication for a biologicalfluid sensor with electronic, electrochemical and chemical components.

The fabrication method comprises steps for manufacturing an apparatuscomprising at least one electrochemical sensor, at least onemicrocontroller, and at least one transceiver. The fabrication processincludes the steps of substrate fabrication, circuit fabrication, pickand place, reflow soldering, electrode fabrication, membranefabrication, sealing and curing, layer bonding, and dressing.

The present invention further includes a metallization paste andsequence step for a reference probe; and a metallization application andsequence step for at least one active probe. The present inventionfurther includes method steps for creating a sensor with line and spacecharacteristics configured and designed for sensing and/or analysis ofsweat flow rate and electrolyte probes. The present invention furtherincludes a method for membrane fabrication with precision ionophoreapplication and curing, and includes a method for dressing fabricationwith laser cutting, bonding, and assembly steps for microfluidic anddressing fabrication.

These and other aspects of the present invention will become apparent tothose skilled in the art after a reading of the following description ofthe preferred embodiment when considered with the drawings, as theysupport the claimed invention.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a chart demonstrating biomarker types, changes in biomarkerlevels during progressive stages of injury, and physiologicalpresentations associated with each stage of injury.

FIG. 2 shows a chart relating sweat rate to concentration ofelectrolytes.

FIG. 3 shows the multiple layers and associated parts of the sensorapparatus and a top perspective view of the complete sensor apparatus.

FIG. 4 shows a perspective top view of a flexible electronic layer onthe bottom adhesive layer.

FIG. 5 shows a complete flexible electronic layer.

FIG. 6 shows a separated layer of electrochemical sensors.

FIG. 7 shows the electrochemical sensors.

FIG. 8 shows a top perspective view (top) and bottom perspective(bottom) view of a complete sensor apparatus.

FIG. 9 shows a diagram of the sweat path through the sensor apparatus.

FIG. 10 shows a diagram of the analytical process within the sensorapparatus.

FIG. 11A shows a table of various sweat characteristics including basicratio for sweat flow rate, body surface area calcs, and sweat loss &body mass loss calcs.

FIG. 11B shows a chart of concentration of ions vs. sweat rate.

FIG. 11C shows tables including work level, zone, SFR, mV ratio, NaLoss, K Loss, Typ Wt, sweating rate and sweat sodium concentration and achart of frequency vs. Na loss.

FIG. 11D shows a table of sweat stds for a typical user.

FIG. 11E shows a chart of oxygen consumption vs. exercise intensity andtables for VO₂ max for men and women.

FIG. 11F shows one embodiment of human use self-calibration for thesensor apparatus.

FIG. 12A shows a table of basic electrolyte concentration conversions.

FIG. 12B shows a table of ratiometric analysis used within the sensorapparatus.

FIG. 12C shows a table of basic mV ratios.

FIG. 13A shows thresholds for electrolyte concentrations.

FIG. 13B shows a table of user input at session start, input from phone,and input from sensor.

FIG. 14 shows a diagram of the sweat sensor subsystem.

FIG. 15 shows a diagram of components within the sweat sensor subsystem.

FIG. 16 shows a diagram of sweat sensor communications.

FIG. 17 shows a diagram of a NXP semiconductor used in an embodiment ofthe sensor apparatus.

FIG. 18 shows a diagram of the communication between a NXP semiconductorand a wireless device in an embodiment of the sensor apparatus.

FIG. 19 shows a diagram of a sweat microcircuit of the sensor apparatus.

FIG. 20 shows a diagram of the system architecture.

FIG. 21 shows a diagram of the controls within the user mobile app ofthe wireless remote transceiver.

FIG. 22 shows a diagram of the network connection between the usermobile app and the user web service.

FIG. 23 shows a diagram of the controls within the cloud database andlibrary, which are part of the user web service.

FIG. 24 shows an individual view of the user web front.

FIG. 25 shows a multi-user view of the user web front.

FIG. 26 shows a diagram of the controls within the present invention'swebsite.

FIG. 27 shows an image of a user data base on the remote computerserver.

FIG. 28 shows a diagram of the generic cloud architecture.

FIG. 29 shows a diagram of a cloud enterprise.

FIG. 30 shows a diagram of another embodiment of a cloud enterprise.

FIG. 31 shows a diagram of a system for epidemiological research.

FIG. 32 shows a diagram of a manufacturing process for the sensorapparatus.

FIG. 33 shows a schematic diagram illustrating general components of acloud-based computer system.

FIG. 34 shows a diagram of a suite of sensors in addition to a sweatsensor.

FIG. 35 shows a top perspective view of one embodiment of the sensorapparatus.

FIG. 36 shows a top perspective view of another embodiment of the sensorapparatus.

FIG. 37A shows a top perspective view of a sensor with a liquidionophore coating.

FIG. 37 B shows a side perspective view of a sensor with a liquidionophore coating.

FIG. 37 C shows another side perspective view of a sensor with a liquidionophore coating.

FIG. 38A shows a top perspective view of a sensor with a liquidionophore coating which was formed using a well approach.

FIG. 38B shows a side perspective view of a sensor with a liquidionophore coating which was formed using a well approach.

FIG. 38C shows another side perspective view of a sensor with a liquidionophore coating which was formed using a well approach.

FIG. 39A shows a top perspective view of a sensor with a liquidionophore coating which was formed using a surface tension dam approach.

FIG. 39B shows a side perspective view of a sensor with a liquidionophore coating which was formed using a surface tension dam approach.

FIG. 40 shows a battery latch mitigation capacitor placement.

FIG. 41 shows an RC configuration with ground reference.

FIG. 42 shows an RC configuration with differential measurement.

FIG. 43 shows an example of cap placement Vcc to Ground for supplyvoltage stability and/or noise immunity.

FIG. 44A shows a side perspective view of a sensor with a liquidionophore coating which was formed using a multilayer substrateapproach.

FIG. 44B shows an exploded side perspective view of the sensor of FIG.44A.

FIG. 45A shows a side perspective view of a sensor with a liquidionophore coating which was formed using another multilayer substrateapproach.

FIG. 45B shows an exploded side perspective view of the sensor of FIG.45A.

FIG. 46A shows a side perspective view of a sensor with an ionophorecovering after a carrier or film is removed following application of apre-formed ionophore to an active electrode.

FIG. 46B shows an ionophore covering on a carrier or film prior toapplication on an active electrode.

FIG. 47 shows a top perspective view, a side perspective view, and anangled perspective view of an ionophore covering formed in a continuousshape that can be handled or applied on a carrier or film that becomesan integral part of the sensor.

FIG. 48 shows a top perspective view, a side perspective view, and anangled perspective view of an ionophore covering formed in anon-continuous shape that can be handled or applied on a carrier or filmthat becomes an integral part of the sensor.

FIG. 49A shows a side perspective view of a sensor with an ionophorecovering applied on a carrier or film that becomes an integral part ofthe sensor.

FIG. 49B shows an ionophore covering on a carrier or film prior toapplication on an active electrode.

FIG. 50 shows a top perspective view and an angled perspective view ofan ionophore formed using an embedded strength approach.

FIG. 51 shows a top perspective view of active and reference electrodesaligned in a linear fashion across the sensor array.

FIG. 52A shows a side perspective view of an ionophore cast and placedon top of an active electrode using an adhesive or bonding film.

FIG. 52B shows an exploded side perspective view of an ionophore castand placed on top of an active electrode using an adhesive or bondingfilm.

FIG. 53A shows a side perspective view of an ionophore cast and placedon top of an active electrode without using an adhesive or bonding film.

FIG. 53B shows an exploded side perspective view of an ionophore castand placed on top of an active electrode without using an adhesive orbonding film.

DETAILED DESCRIPTION

The present invention includes methods of fabrication or manufacturing abiological fluid sensor device with electronic, electrochemical andchemical components for sensing and collecting biological fluidbiomarkers, transmitting data to a transceiver device on a remotecomputing device for calculating biomarker data, and analyzing the data,and storing the data on the remote computing device or on a remotecomputer server.

The present invention provides methods of making the sensor apparatus ordevice for sensing sweat biomarkers that are operable and functional asdescribed herein. A preferred embodiment is surface mount technology; itmay provide better electrical performance compared to hand soldering; itallows use of smaller electrical components, denser layout, cheaperboards, and improved shock and vibration characteristics. Electricalcomponents include chips, resistors, transistors, and capacitors. Anautomated dispenser configuration for dispensing an ionophore ispreferably used to ensure a higher unit production rate when comparedwith the prior art. The automated dispenser provides for precise andrapid ionophore coating.

The sequence of the steps as well as the steps themselves is critical tothe operational use and accuracy of the sensor apparatus. It is criticalto initially install the surface mount parts and run through solderreflow before assembly of the sensor units. Subsequently, the heatsensitive parts to include the reference electrode, active electrodes,and ionophore coatings should be applied in that order. Following thelast ionophore coating, the entire flexcircuit layer needs a minimumabout 3 hour cure, which can be accelerated by UV and/or other rapidheating/drying processes. Following the cure period, the sensors can befinal-assembled with the medical top cover, microfluidic, anddouble-sided skin adhesive layers using an industrial grade epoxy.

In a preferred embodiment, a microcircuit is printed directly on theflexible substrate using bare die bonding. The microcircuit is attachedto the flexible substrate using any common bare die bonding process,such as eutectic, solder, adhesive, glass or silver-glass.

The fabrication process includes the steps of substrate fabrication,circuit fabrication, pick and place, reflow soldering, electrodefabrication where the earlier heat intensive processes through refloware completed prior to the heat sensitive processes associated withstarting with the build up and treatment of metalized traces on thesubstrate followed by membrane fabrication, sealing and curing, anddressing. In one embodiment, the substrate is a Kapton substrate.Without this process sequence, the electrodes build up unpredictablelevels of oxidation and other undesirable surface conditions whichnegatively affect the accuracy within a sensor, as well as amongmultiple sensors. In a similar manner, the heat intensive steps alsodegrade/melt the ionophore coating, a polymer not tolerant of elevatedtemperature. Repeated heating and cooling exposes the ionophore tochemical changes and also creates microscopic holes in the coating, bothwhich allow untargeted biomarker ions to penetrate the coating to reachthe electrode which generates potential strength not associated with thetarget biomarker, thus reducing the accuracy.

The present invention further includes a metallization paste made of astable metal (silver, gold, platinum, palladium) on top of the circuittrace metal (typically copper or gold) which is further stabilizedthrough chemical and/or thermal annealing treatments to construct areference probe; and a similar metallization application and sequencestep for at least one active probe. The present invention furtherincludes method steps for creating a sensor with line and spacecharacteristics configured and designed for sensing and/or analysis ofsweat flow rate and small protein probes.

The present invention further includes a method for membrane fabricationwith precision ionophore application and curing. Sensor functionalityand accuracy require precision placement with proper thickness of asmall amount of ionophore polymer on the active electrodes tofilter/prevent untargeted ions to reach the electrode. The amount isapproximately 2 microliters with a designated viscosity placed in aclean assembly environment to completely cover the exposed activeelectrode on the skin-facing side of the flexcircuit. The coating shallpreferably not exceed more than 0.5 mm from the edge of the electrode.The placement also preferably includes automated dispenser configurationand settings to ensure high speed or high rate manufacturing as well asmanufacturing consistency that minimizes manufacturing accuracyvariations.

The present invention still further includes a method for dressingfabrication with laser cutting, bonding, and assembly steps formicrofluidic and dressing fabrication. Following the designatedionophore cure time, the flexcircuits with integrated parts, sensorelectrodes are conformal sealed and cured. The sealed flex circuits areplaced on a large sheet of top cover medical textile with previouslylaser precut patterns and adhesive up. The sealed flexcircuits adhere tothe top cover via the cover's adhesive; they are mechanicallyprecision-placed and pressed into place with the exposed sensor headfacing up for permanent bonding using proprietary edge bonding patternsto allow for the proper flow/exit of sweat from the sensor heads out.Next, microfluidic units previously pre-cut to specification areinserted on top via the same mechanical precision placement equipment.And finally the mechanical placement equipment places a large sheet ofdouble sided adhesive which were previously precut by laser to the samesize and pattern as the top cover textile. The sheet and each of thelaser precut patterns are precisely positioned above the semi-assembledunits below and pressed into position for permanent bond. The individualunits are then mechanically separated (punched) from their host sheetsand collected in bins for automated packaging into individualizedbranded wrappers, and then inserted into branded boxes that are cratedand palletized for shipment. The wrapper, box, crate, and palletspecifications are specified by our distribution partners' pack out planand specified branding schema.

FIG. 3 illustrates a multi-layered apparatus or device that includes adouble-sided adhesive layer, a filter paper, an electronic layer, and awoven top adhesive. More specifically, the device is flexible andmulti-layered, wherein the layers comprise the following: amacrofluidic, double-sided adhesive layer; an electronic layercomprising at least one electrochemical sensor, a microcontroller, and atransceiver antenna coil; a microfluidic management layer; and a vaporporous, top protective layer. The macrofluidic, double-sided adhesivelayer is intimately adhered to the skin. The electronic layer isintimately adhered to the microfluidic, double-sided adhesive layer, asshown in FIG. 4 . The microfluidic management layer circumferentiallysurrounds the at least one electrochemical sensor of the electroniclayer. The vapor porous, top protective layer is placed on andcompletely covers the microfluidic management layer and electroniclayer. The vapor-porous, top protective layer is intimately adhered tothe macrofluidic, double-sided adhesive layer. The fully fabricatedsensor apparatus is shown in FIG. 8 . Preferably, the length of theapparatus is approximately 76.1 mm. In one embodiment, the adhesive ofthe apparatus is Double Coated Polyester Nonwoven Tape (commerciallyavailable as 3M 9917 as of this writing). In another embodiment, theadhesive of the apparatus is Tan Tricot Knit Tape (commerciallyavailable as 3M 9926T as of this writing). The design of themicrofluidic layer improves flow control and decreases patch layerdelamination during high sweat volume use cases.

FIG. 34 shows a diagram illustrating a suite of sensors in addition to asweat sensor, including a non-invasive penetration (NIP) sensor, anaudio sensor, a vitals sensor, an electro-optical/infrared sensor, anoxygen sensor, a breath sensor, and a sweat sensor. In one embodimentthe sensor includes at least two of the sensors in FIG. 34 . In anotherembodiment, one of the suite of sensors is the only sensor utilized. Oneor more of the sensors is embedded in the skin in one embodiment of thepresent invention. An embedded sensor preferably mimics the compositionand behavior of cells. The electro-optical/infrared sensor may include afluorescent signal sensor. In one embodiment, a reader sends anexcitation light through the skin to the biosensor, which then emits afluorescent light proportional to the amount of biochemical measured.

FIG. 35 shows a top perspective view of one embodiment of the sensorapparatus. The flex circuit is substantially centrally located on thebandage material and adhesive. Wicking paper is utilized to move bodilyfluid, particularly sweat, through the sensor apparatus. The reverse “D”shaped hole is through the adhesive and the wicking paper. Alldimensions in FIG. 35 are in MM or VOS, as appropriate.

FIG. 36 shows a top perspective view of another embodiment of the sensorapparatus. The flex circuit is substantially centrally located on thebandage material and adhesive. Wicking paper is utilized to move bodilyfluid, particularly sweat, through the sensor apparatus. The reverse “D”shaped hole is through the adhesive and the wicking paper. Alldimensions in FIG. 36 are in MN/I or VOS, as appropriate.

The sensor apparatus is designed to allow sweat to flow through lasercut, macrofluidic pores in the skin adhesive layer, as shown in FIG. 9 .Sweat then flows through a filter to the electronics layer, specificallythe electrochemical sensor unit, where biomarkers may contact theelectrodes of the electrochemical sensor unit. The sweat evaporatesthrough the woven textile protective top layer. The evaporation affordsimproved and continuous sweat flow into the sensor apparatus. Thiswicking ensures sweat sensing measures are consistently using new sweatsamples rather than static or diluted samples. In one embodiment, thewicking and sweat flow rates range from 0% to 5% of Total Body Loss/hr(instant equivalent sweat loss rate) or equivalent to 11.5 L/hr totalloss.

The present invention further includes a device with a small amount ofionophore polymer on the active electrodes to filter/prevent untargetedions to reach the electrode. Sensor functionality and accuracy requireprecision placement with proper thickness of a small amount of ionophorepolymer in one embodiment. The precision placement is conducted eithermanually or via automated equipment. The amount of ionophore polymer isapproximately 2 microliters with a designated viscosity placed in aclean assembly environment to completely cover the exposed activeelectrode on the skin-facing side of the flexcircuit. The coating shallpreferably not exceed more than 0.5 mm from the edge of the electrode.In one embodiment, the ionophore polymer is cured. In anotherembodiment, the curing takes place using heat and/or light to acceleratedrying without changing the ionophore selectivity characteristics.

In one embodiment, the sensor is calibrated. Preferably, a human usercalibrates the sensor using actual test results and feedback from thesensor. Advantageously, human user calibration or human useself-calibration (H-SCAL) provides for more accurate data when comparedwith calibration of sensors solely in the laboratory. Actual human testdata is compared to external sweat loss values obtained from highlyaccurate scales. The external data is utilized to adjust several factorsfor a human user. Specifically, the adjustments provide for correctionof a collection of diminutive and/or major error sources to improve theoverall accuracy of the system.

FIG. 11F details one embodiment of the H-SCAL wherein the apparatus iscalibrated using a personalization factor (Fp). The default value of thepersonalization factor is 1. At least one remote transceiver devicecalculates the personalization factor using at least a weight of theuser before physical activity and a weight of the user after physicalactivity. In a preferred embodiment, the weight of the user beforephysical activity and the weight of the user after physical activity aremanually entered into the remote transceiver device. In an alternativeembodiment, the weight of the user before physical activity and theweight of the user after physical activity are automatically uploaded bya smart scale and/or a third party application. The at least one remotetransceiver device transmits the personalization factor to at least oneremote computer server or at least one remote computing device ordatabase for storage. The personalization factor is updated when a newweight of the user before physical activity and a new weight of the userafter physical activity are provided. In one embodiment, thepersonalization factor is further based on a maximum rate of oxygenconsumption for the human (VO₂ max), fitness or conditioning level, abody mass factor, outdoor temperature, humidity, and/or estimated bodysurface area. In one embodiment, the personalization factor is used toadjust the values of Fcondition and/or SFR. In an alternativeembodiment, the value of Fcondition varies depending on the value of VO₂max. The personalization factor corrects for normal variations in ahuman from time to time, and also normalizes minor variations due tomanufacturing, storage, wetting, and handling.

The sensor apparatus includes sweat sensor subsystem, as shown in FIG.15 , which includes a microcontroller that receives multiple input data,which are input from multiple sources. A first source is biologicalfluid, preferably sweat, although alternative fluids may be used. Thesweat contains a variety of analytes, such as, by way of example and notlimitation, electrolytes, small molecules (molecular weight <900Daltons), proteins, and metabolites. Exemplary analytes includesubstances including sodium or potassium. The article The microfluidicsof the eccrine sweat gland, including biomarker partitioning, transport,and biosensing implications, by Sonner, et al in Biomicrofluidics 2015May; 9(3); 031301, which is incorporated by reference herein in itsentirety, reports microfluidic models for eccrine sweat generation andflow and reviews blood-to-sweat biomarker partition pathways. In oneembodiment, the sensor apparatus is operable to sense sodium andchloride both in a dynamic range from about 0 mM to about 120 mM, withnormal ranges in humans being about 20 mM to about 100 mM. In anotherembodiment, the sensor apparatus is operable to sense potassium in adynamic range from about 0 mM to about 40 mM, with normal ranges inhumans being about 5 mM to about 20 mM. In one embodiment, the sensorhas a response time of about 60 seconds with about a 90% response. Otheranalytes include oxygen, glucose, calcium, ammonium, copper, magnesium,iron, zinc, lactate, creatinine, uric acid, urea, ethanol, amino acids,hormones, steroids, proteins, catecholamines, and interleukins. In oneembodiment, the sensor is operable to analyze the analytes at a pMlevel, preferably in the 1-10 pM range or even below 1 pM (the sub-pMlevel). These analytes are collected at the electrochemical sensor, asshown in FIG. 7 , which houses reference (preferably standard) andactive electrodes, wherein, by example and not limitation, theelectrodes are silver, zinc, copper, gold, platinum, rhodium, carbon ora combination thereof. In one embodiment, the apparatus has an embeddeddot-circle configuration for a reference electrode to improve stabilitythrough less interference. Additionally, gold probes or electrodes areused in one embodiment to improve stability and reduce production costs.The apparatus also includes a microprocessor, multiplexer (mux), ADC,and optimized on board processing for real time, pre-transmission sensorsignal conditioning in another embodiment.

Unique human induced electromagnetic interference (H-EMI) sometimescause interference and produce inaccurate measurements from the sensorapparatus. Specifically, human induced anomalies affect flex circuitfunctionality, performance, and reliability. Variations in the locationof the sensor on the human body as well as human skin variations betweenpeople can cause unpredictable flex circuit behavior that is not readilyapparent in lab settings. Through testing on humans, various embodimentsand solutions to H-EMI have been developed. In one embodiment, hardwarecomponents are placed to mitigate the human use electromagneticinterference effects on flex circuits. Specifically, the placement ofcapacitors compensates for intermittent power variations. Additionally,strategically placed Kapton reinforcements (or other polyimidecomponents) further mitigate EMI disturbances resulting from human useelectromagnetic interference. Specifically, capacitors are utilized onpower/battery latching circuits to mitigate human motion artifactsimpacting measurement cycles. FIG. 40 shows a battery latch mitigationcapacitor placement. The values included in FIG. 40 are for purposes ofillustration by example and in no way limit the values which areutilized in the present invention.

Adjustments in firmware running on a microcontroller also offset EMI insome embodiments. In another embodiment, adjustments to filtering(preferably by adding specific RLC components) and noise suppressionoffset EMI. Improved measurement electronics input design and samplingmethods also mitigate H-EMI. In one embodiment, input design involvescomplex impedance related to the RLC (mostly R and C) components tocondition sensor inputs to overcome motion artifacts manifest from thehuman sensor interface and input buffering methods to reduce measurementelectronics impact on sensor measurements (Heisenberg Uncertainty). FIG.41 shows an RC configuration with ground reference. FIG. 42 shows an RCconfiguration with differential measurement. FIG. 43 shows an example ofcap placement Vcc to Ground for supply voltage stability and/or noiseimmunity. Firmware branching logic preferably differentiates betweenpower on cycle and motion induced power variations.

The present invention also includes sensor embodiments which areoperable under the most demanding physical environments, and inparticular, athletic use cases. Specifically, a durable sensor is neededfor these cases because of exposure to violent impact shock, speedchanges, motion intensity, exposure to water, etc. Strengthening theflex circuit and electronic layouts minimize use case impacts on thesensor. Additionally, advanced flex circuit protection minimizes impactsupon the sensor. Advanced flex circuit protection includes strategicflex circuit mix with rigid boards and/or application-specificintegrated circuit (ASIC) miniaturization. Advanced rubberized casingsand microfluidics based on crystal fiber technology are also utilized inone embodiment. Specifically, the present invention includes theaforementioned adjustments to the sensor head of the sensor apparatus.Sensor heads accommodate a variety of motion factors, including flex,stretch, sliding, and shock loading. In one embodiment, the presentinvention utilizes multiple sensor configurations to accommodateindividual sensor disruption and/or failure.

In one embodiment, the sensor head is detachable and reattachable. Thesensor head is attached via z-axis tape or hot bar soldering in oneembodiment. In another embodiment, there is a standardizedinterconnection. In yet another embodiment, the sensor head pinout isselectable from the multiplexer side. In one embodiment, uC and a RFantenna are integrated with fabric. In yet another embodiment, two partencryption is utilized. RFI and shielding as well as adding layers tothe board to provide additional ground planes and/or metallized fabricin dressing are also utilized.

Additionally, flexibility in most components of the sensor apparatus isnot desired. FR4 is utilized in a preferred embodiment. The sensor headis the only flexible portion of the sensor in one embodiment, as thesensor head requires flex and adhesion to the human. However, forembodiments in which flexibility is desired, a variety of flexiblesubstrates are utilized.

Another embodiment of the present invention includes adding a layer orrow of 0 ohm resistors on either side of the multiplexer. Additionally,buffer amps are utilized either before or after the multiplexer. In analternative embodiment, higher impedance ADC is utilized. Faultdetection and isolation is also used in the systems and methods of thepresent invention.

FIG. 37A shows a top perspective view of a sensor with a liquidionophore coating. A circuit board or other substrate 701 includes acopper trace or other conductive material/circuit element 703, anionophore or any material applied using a liquid deposition method 705,and copper or other conductive material forming an active electrode 707.The ionophore 705 preferably covers the copper or other conductivematerial forming an active electrode 707.

FIG. 37 B shows a side perspective view of a sensor with a liquidionophore coating.

FIG. 37 C shows another side perspective view of a sensor with a liquidionophore coating.

One sensor head embodiment of the present invention includes aRing-Reference design. This design preferably solves issues with surfacetension management.

FIG. 39A shows a top perspective view of a sensor with a liquidionophore coating which was formed using a surface tension dam approachor Ring-Reference design. A circuit board or other substrate 701includes a copper trace or other conductive material/circuit element703, an ionophore or any material applied using a liquid depositionmethod 705, copper or other conductive material forming an activeelectrode 707, and a soldermask, printed ink, or any othernon-conductive material dissimilar to the circuit board printed,deposited to, or otherwise adhered to the circuit board prior to liquiddeposition 709. The ionophore 705 preferably covers the copper or otherconductive material forming an active electrode 707.

FIG. 39B shows a side perspective view of a sensor with a liquidionophore coating which was formed using a surface tension dam approachor Ring-Reference design.

In another embodiment, a surface tension management solution includes aprinted or screened non-conductive ring inside a ring electrode tocreate a surface tension discontinuity. A soldermask preferably is asurface tension discontinuity in the surface tension managementsolution. In another embodiment, printed ink is a surface tensiondiscontinuity in the surface tension management solution. Printed inkrefers to any printed, screened, or deposited non-conductive material.This surface tension discontinuity supports the uniform deposition ofliquid ionophores. In another embodiment, a well design is utilized forthe sensor head for ionophore and electrode isolation. Specifically, thewell design mitigates the stretch and/or sliding issues by increasingabrasion resistance.

FIG. 38A shows a top perspective view of a sensor with a liquidionophore coating which was formed using a well approach. A circuitboard or other substrate 701 includes a copper trace or other conductivematerial/circuit element 703, an ionophore or any material applied usinga liquid deposition method 705, and copper or other conductive materialforming an active electrode 707. The ionophore 705 preferably covers thecopper or other conductive material forming the active electrode 707.

FIG. 38B shows a side perspective view of a sensor with a liquidionophore coating which was formed using a well approach.

FIG. 38C shows another side perspective view of a sensor with a liquidionophore coating which was formed using a well approach.

The Ring-Reference design or surface tension management solution and thewell design are used to manage ionophore deposition, specifically liquiddeposition on a surface. The liquid is preferably deposited on thesurface of a circuit board and then solidifies and/or hardens.

Roll-to-roll manufacturing is an example of a manufacturing method thatis efficient, cost effective, and allows for high production rates.Conventional processing (e.g., batch processing) is slower and highercost due to the numerous steps involved. For example, liquid depositionof the ionophore is costly. Alternative methods, such as roll-to-rollmanufacturing, may also improve the reliability of the system. However,roll-to-roll manufacturing requires a more durable ionophore polymercoating. One method of manufacturing ISEs with ionophore coverings thatcould be used with roll-to-roll manufacturing is to pre-fabricate theionophore covering. In one embodiment, a pre-formed ionophore coveringis adhered to the active electrode as an alternative to direct liquiddeposition. In another embodiment, a durable ionophore layer ispre-fabricated and applied via a mechanical method. In yet anotherembodiment, a pre-formed ionophore covering is attached by an adhesivemeans (e.g., conductive adhesive) or thermal reflow.

A pre-formed ionophore covering can be created using a commerciallyavailable ion selective material dissolved in a polyvinyl chloride (PVC)solution. This requires a method of forming the pre-formed ionophorethat increases the durability of the resulting ‘plastic’ ionophorecovering. Various forming methods are employed to facilitate theplacement of the ionophore covering on the active electrode. Forexample, the ionophore is cast in a shape and placed on a carrier or afilm that facilitates mechanical placement via a pick-and-place orsimilar device manufacturing process. A cast shape requires a boundingform or structure that assures shape and handling consistency. This castshape requires a means or method of adhesion applied locally to theactive electrode without damaging or deforming the ionophore covering.

One such approach is to separate the active and reference electrodesonto different substrate layers (“a multilayer substrate approach”). Theat least one reference electrode is on a first substrate layer that hasvoids or wells into which the ionophore is cast or formed and retainedwithin the first substrate layer. The first substrate layer is thenbonded in its entirety to a second substrate layer containing the atleast one active electrode properly aligned with the wells containingthe formed ionophore.

FIG. 44A shows a side perspective view of a sensor with a liquidionophore coating which was formed using a multilayer substrateapproach. A copper or other conductive material forms an activeelectrode 707. The active electrode is on top of an active electrodesubstrate 717. An adhesive or bonding film 715 is deposited on top ofthe active electrode substrate 717. Reference electrodes 711 are on topof a reference electrode substrate 713. An ionophore is cast in a wellof the reference electrode substrate 713 to create a pre-formedionophore 719. The reference electrode substrate 713 containing thepre-formed ionophore 719 is attached to the active electrode substrate717 using the adhesive or bonding film 715. In a preferred embodiment,the adhesive or bonding film is a conductive adhesive and the referenceelectrode substrate and active electrode substrate are non-conductive.In a preferred embodiment, the copper or other conductive material issilver and the reference electrode substrate and active electrodesubstrate are comprised of polyester. The pre-formed ionophore requiresthermal reflow because the active electrode is not covered with aconductive adhesive.

FIG. 44B shows an exploded side perspective view of the sensor of FIG.44A.

FIG. 45A shows a side perspective view of a sensor with a liquidionophore coating which was formed using another multilayer substrateapproach. A copper or other conductive material forms an activeelectrode 707. The active electrode is on top of an active electrodesubstrate 717. An adhesive or bonding film 715 is deposited on top ofthe active electrode substrate 717 and the active electrode 707.Reference electrodes 711 are on top of a reference electrode substrate713. An ionophore is cast in a well of the reference electrode substrate713 to create a pre-formed ionophore 719. The reference electrodesubstrate 713 containing the pre-formed ionophore 719 is attached to theactive electrode substrate 717 using the adhesive or bonding film 715.In a preferred embodiment, the adhesive or bonding film is a conductiveadhesive and the reference electrode substrate and active electrodesubstrate are non-conductive. In a preferred embodiment, the copper orother conductive material is silver and the reference electrodesubstrate and active electrode substrate are comprised of polyester. Thepre-formed ionophore requires thermal reflow when the active electrodeis not covered with a conductive adhesive.

FIG. 45B shows an exploded side perspective view of the sensor of FIG.45A.

Alternatively, the ionophore covering is formed in a shape that can behandled or applied on a carrier or film that can be removed afterapplication. In another embodiment, the ionophore covering is formed ina shape that can be handled or applied on a carrier or film that becomesan integral part of the sensor (e.g., a microfluidic absorption layer).

FIG. 46A shows a side perspective view of a sensor with an ionophorecovering after a carrier or film is removed following application of apre-formed ionophore to an active electrode. A copper or otherconductive material forms an active electrode 707. The active electrode707 and reference electrodes 711 are positioned on top of a substrate701. An ionophore is cast to create a pre-formed ionophore 719 that isattached to a carrier or film (not shown). The pre-formed ionophore 719and the carrier or film (not shown) is placed on top of the activeelectrode 707 using an adhesive or bonding film 715. The carrier or film(not shown) is then removed. In a preferred embodiment, the adhesive orbonding film is a conductive adhesive and the substrate isnon-conductive. In a preferred embodiment, the copper or otherconductive material is silver and the substrate is comprised ofpolyester. The adhesive or bonding film does not contact adjacentelectrodes in a preferred embodiment. Alternatively, a z-axis tape isused to prevent adjacent electrodes from shorting.

FIG. 46B shows an ionophore covering on a carrier or film prior toapplication on an active electrode. The pre-formed ionophore 719 isattached to the carrier or film 721. The pre-formed ionophore 719 andthe carrier or film 721 are placed on top of the active electrode 707using the adhesive or bonding film 715. The carrier or film 721 is thenremoved from the pre-formed ionophore 719.

FIG. 47 shows a top perspective view, a side perspective view, and anangled perspective view of an ionophore covering formed in a continuousshape that can be handled or applied on a carrier or film that becomesan integral part of the sensor. An ionophore is cast to create apre-formed ionophore 719. The pre-formed ionophore 719 is attached to acarrier or film 721. The pre-formed ionophore 719 and the carrier orfilm 721 are placed on top of an active electrode (not shown) using anadhesive or bonding film 715. In a preferred embodiment, the adhesive orbonding film is a conductive adhesive. In a preferred embodiment, thecopper or other conductive material is silver and the substrate iscomprised of polyester. The adhesive or bonding film does not contactadjacent electrodes in a preferred embodiment. Alternatively, a z-axistape is used to prevent adjacent electrodes from shorting. In anotherembodiment, the carrier or film acts as a microfluidic absorption layer.The carrier or film is microfluidic paper in one embodiment. In yetanother embodiment, the pre-formed ionophore is attached to the carrieror film using a cast bond or an adhesive.

FIG. 48 shows a top perspective view, a side perspective view, and anangled perspective view of an ionophore covering formed in anon-continuous shape that can be handled or applied on a carrier or filmthat becomes an integral part of the sensor. An ionophore is cast tocreate a pre-formed ionophore 719. The pre-formed ionophore 719 isattached to a carrier or film 721. The pre-formed ionophore 719 isseparated on either side by voids 725. The pre-formed ionophore 719 andthe carrier or film 721 are placed on top of an active electrode (notshown) using an adhesive or bonding film 715. In a preferred embodiment,the adhesive or bonding film is a conductive adhesive. In anotherembodiment, the carrier or film acts as a microfluidic absorption layer.The carrier or film is microfluidic paper in one embodiment. In yetanother embodiment, the pre-formed ionophore is attached to the carrieror film using a cast bond or an adhesive.

FIG. 49A shows a side perspective view of a sensor with an ionophorecovering applied on a carrier or film that becomes an integral part ofthe sensor. A copper or other conductive material forms an activeelectrode 707. The active electrode 707 and reference electrodes 711 arepositioned on top of a substrate 701. An ionophore is cast to create apre-formed ionophore 719. The pre-formed ionophore 719 is attached to acarrier or film 721. The pre-formed ionophore 719 and the carrier orfilm 721 are placed on top of the active electrode 707 using an adhesiveor bonding film 715. In a preferred embodiment, the adhesive or bondingfilm is a conductive adhesive. In a preferred embodiment, the copper orother conductive material is silver and the substrate is comprised ofpolyester. The adhesive or bonding film does not contact adjacentelectrodes in a preferred embodiment. Alternatively, a z-axis tape isused to prevent adjacent electrodes from shorting. In anotherembodiment, the carrier or film acts as a microfluidic absorption layer.The carrier or film is microfluidic paper in one embodiment. In yetanother embodiment, the pre-formed ionophore is attached to the carrieror film using a cast bond or an adhesive.

FIG. 49B shows an ionophore covering on a carrier or film prior toapplication on an active electrode.

In another embodiment, the ionophore is formed with an embedded strengthmember or structure that does not impair the function of the ionophore.For example, the ionophore is cast with a glass or plastic fiberembedded in the material so that it is more durable and can be handledwith greater ease in various manufacturing processes. This is similar inprinciple to rebar in concrete. This embedded strength approach permitslaser cutting of shapes for pick-and-place manufacturing or reel-typedispensing for inline or roll-to-roll manufacturing methods.

FIG. 50 shows a top perspective view and an angled perspective view ofan ionophore formed using an embedded strength approach. An ionophore iscast to create a pre-formed ionophore 719 with an embedded glass orplastic fiber 727. A cubic shape is shown in the figure, but othershapes (e.g., hexagonal, tetragonal, orthorhombic, rhombohedral,triclinic, monoclinic) are compatible with the present invention.

In one embodiment, the active and reference electrodes are aligned in alinear fashion across the sensor array to permit continuous placement ofa formed or cast ionophore as a ‘tape’ for roll-to-roll processing. Aspreviously mentioned, roll-to-roll manufacturing is efficient, costeffective, and allows for high production rates.

FIG. 51 shows a top perspective view of active and reference electrodesaligned in a linear fashion across the sensor array. A copper or otherconductive material forms active electrodes 707. Reference electrodes711 are aligned in a linear fashion across the sensor array. Anionophore is cast to create a pre-formed ionophore 719 and placed on topof the active electrodes 707 using an adhesive or bonding film 715. In apreferred embodiment, the adhesive or bonding film is a conductiveadhesive. In a preferred embodiment, the copper or other conductivematerial is silver and the substrate is comprised of polyester. Theadhesive or bonding film does not contact adjacent electrodes in apreferred embodiment. Alternatively, a z-axis tape is used to preventadjacent electrodes from shorting.

FIG. 52A shows a side perspective view of an ionophore cast and placedon top of an active electrode using an adhesive or bonding film. Acopper or other conductive material forms an active electrode 707. Theactive electrode 707 and reference electrodes 711 are on top of asubstrate 701. An ionophore is cast to create a pre-formed ionophore719. The pre-formed ionophore is placed on top of the active electrode707 using an adhesive or bonding film 715. In a preferred embodiment,the adhesive or bonding film is a conductive adhesive. In a preferredembodiment, the copper or other conductive material is silver and thesubstrate is comprised of polyester. The adhesive or bonding film doesnot contact adjacent electrodes in a preferred embodiment.Alternatively, a z-axis tape is used to prevent adjacent electrodes fromshorting.

FIG. 52B shows an exploded side perspective view of an ionophore castand placed on top of an active electrode using an adhesive or bondingfilm.

FIG. 53A shows a side perspective view of an ionophore cast and placedon top of an active electrode without using an adhesive or bonding film.A copper or other conductive material forms an active electrode 707. Theactive electrode 707 and reference electrodes 711 are on top of asubstrate 701. An ionophore is cast to create a pre-formed ionophore719. The pre-formed ionophore 719 is placed on top of the activeelectrode 707. In a preferred embodiment, the copper or other conductivematerial is silver and the substrate is comprised of polyester. Thepre-formed ionophore requires thermal reflow because the activeelectrode is not covered with a conductive adhesive.

FIG. 53B shows an exploded side perspective view of an ionophore castand placed on top of an active electrode without using an adhesive orbonding film.

Other variations in inter-sensor spacing are also utilized in thepresent invention. Variations in inter-sensor spacing improve sensorperformance on a human-sensor interface. Specifically, closer spacingbetween sensors generally contributes to crosstalk and parasitics. Thelocation and proximity of a reference sensor can also prevent issues forchemical, electrical, and mechanical isolation. Both chemical andmechanical interactions can compromise measurements of adjacent sensors,especially when sensors are of dissimilar construction and/or electricalspecification. Preferably, the location of a chloride sensor relative toa sodium sensor and/or a potassium sensor is varied to increase abrasionresistance, which mitigates stretching and/or sliding issues.Preferably, the location of the chloride sensor is not proximal to thelocation of Na and K sensors to avoid chemical and mechanicalinteractions between the sensors.

The sensor also includes a galvanic skin response sensor in oneembodiment. A longer lead and/or additional ground plane under thebattery are preferably utilized to resolve complications resulting fromadding a galvanic skin response sensor and/or a chlorine sensor.

In yet another embodiment, utilizing alternative flex substrates andconductor construction provides increase abrasion resistance.Preferably, silver ink on polyester or a material similar to polyesteris utilized.

Preferably, the sensor of the present invention does not include ajumper or a Galvanic Skin Response (GSR). In one embodiment, FR4 isadded under molex or directly integrated into the BR/uC and RF antenna.

The apparatus is also optimized for efficient and successfultransmission during athletic usage. When the electrodes contact thesweat biomarkers a voltage is produced. Three electrodes per analyte areused to create an average voltage value, which is transmitted to themicrocontroller, wherein the microcontroller pre-processes and preparesthe sensor data to be communicated to the transceiver device viaBluetooth for providing near field communication in preferredembodiments. Alternatively, an RFID, NFC, or other proprietarycommunications chip may be provided. The NFC chip preferably has anincreased base signal amplitude for better processing and lowerresolution as well as better concentration confidence and resolution.Bluetooth is preferred due to its low energy, ubiquity, and low cost.Most any Bluetooth enabled device can pair with another Bluetooth devicewithin a given proximity, which affords more ubiquitous communicationbetween the microcontroller and a transceiver device. The dynamic andautomatic connections of Bluetooth allow for multiple microcontrollersto communicate with a single transceiver device, which, by way ofexample and not limitation, would provide for a team-based situation,wherein the sweat data of multiple athletes is communicated to a singlecoach or team database.

Another embodiment provides for utilizing NFC and/or onboard power forsystem control and operation, NFC and serial interfaces for datatransport, external range extenders, and system integration.

Preferably, the systems and methods of the present invention are sensoragnostic, meaning that the systems and methods work with a variety ofsensors. By way of example and not limitation, the systems and methodsof the present invention work with multiple sensor head configurations,including variations in sensor count, single reference electrodesensors, multiple reference electrode sensors, a variety of analyteconcentration, a variety of analyte sensitivity, a variety of inputimpedances, analog measurement conditioning, digital sampling, etc.Notably, variations in hardware and/or firmware designs provide for thesensor agnostic systems and methods of the present invention. Anexemplary hardware implementation of a configurable sensor interface formultiple pinout permutations and variable analog buffering/signalconditioning supports existing and future sensor designs. Exemplaryfirmware designs for sensitivity and noise mitigation include, but arenot limited to, variable input impedance, sampling intervals, settlingtime, and input switching designs. Additionally, addling a settlingdelay between readings also mitigates noise. One embodiment of asettling delay includes switching the multiplexer, waiting 10 ms, thentaking a reading. Another embodiment for noise mitigation includeslowering the gain on the analog digital converter (ADC), which raisesinput impedance, to produce higher voltage levels. Adding both asettling delay and a lower ADC gain together significantly mitigatenoise.

Utilizing non-adjacent channel switching on a multiplexer also reducesnoise in the form of crosstalk and/or ghosting. Specifically, firmwaremethods for sensor switching and measurement times include non-adjacentmultiplexer selection and sensor specific settling times from sensorselection to ADC sampling. Standard single chip multiplexers canexperience adjacent channel crosstalk or ‘ghosting’ from large impedancechanges that can manifest as noise or erroneous measurements. Firmwaresolutions to avoid direct adjacency in measurement selection can reducethese effects and improve measurement electronics performance. This isparticularly important in low signal environments.

Complex sensor selection methods can propagate transition noise into themeasurement electronics. The settling time constant for different sensortypes and variations due to sensor-human interactions can present widefluctuations that are hard to manage with filtering. Firmware controlledselection-to-measurement time delays can mitigate these effects. Thiscan be implemented on a per sensor type basis for systems withdissimilar sensor types. As an example, a ‘ring reference’ ion-selectiveelectrode (ISE) has more localized/uniform human contact behavior acrossthe active sensor and its associated reference. In contrast, a ‘singlereference-multiple sensor’ ISE can have widely differing human contactbehavior due to the distributed physical placement of the singlereference and the specific sensor heads.

The systems and methods of the present invention preferably enableend-to-end flow and processing using a patch, wherein the data iseventually transmitted to a device or to the cloud for data processingand analytics. The patch preferably includes electronics and firmware toproperly buffer, amplify, and manage timing required to optimize patchfunctions. The firmware is preferably modularized to enable engineers toset designated variables, filters, noise thresholds, and otherattributes needed to accommodate many different sensor types,modalities, sensitivities, and other characteristics.

The present invention also provides systems and methods for addressingfault detection and isolation, electromagnetic compatibility (EMC)detection, radiofrequency interference detection, mitigation and eventhandling, addition of encryption for data integrity, personallyidentifiable information (PII) protection, and communications security.

A second source of input data is the remote transceiver device. Bytwo-way communication, the transceiver device may transmit data to themicrocontroller of the apparatus, which is part of an inter-integratedcircuit, as shown in FIGS. 17 and 18 . The data to be transmitted willhave been manually or automatically input in the transceiver device. Forexample and not limitation, as shown in FIG. 10 , types of manuallyinput data may include gender, fitness or conditioning level, age, andanthropomorphic data such as height and weight. The anthropomorphic datais preferably used to estimate user body surface area, which is acritical variable for accurately determining sweat loss and electrolyteloss. More preferably, estimates are a product of anthropomorphic data,gender, and age. Prior art assumes a body surface area of about 2 m² tocalculate sweat loss and electrolyte loss. Using anthropomorphicvariables, as in the present invention, consistently decreasescalculated error rate from between about 50 and about 70 percent to lessthan about 10 percent, preferably. The accuracy resulting from body massestimation revealed that persons with larger body mass, such as males,more readily adapt to physical exertion by sweating more quickly, alarger volume, and lower electrolyte concentration. Similarly, aphysically fit person with a small body mass, such as a female, adaptsto physical exertion by adjusting sweat flow rates and electrolytelevels. Although prior art has validly analyzed sodium, potassium, sweatrates, etc., it has failed to account for body surface area, mass andVO₂ max (maximum rate of oxygen consumption), thus inflating calculatederror rate. These data support that sweat flow rates and electrolyteloss is strongly correlated with body size and surface area andconditioning level, which further supports the need for properestimation of body size, such as through anthropomorphic variables.

Accurately measuring VO₂ max generally requires testing with a treadmillor bicycle in a laboratory. Exercise intensity is progressivelyincreased while measuring the volume of inhaled and exhaled air and theoxygen and carbon dioxide concentrations of inhaled and exhaled air. VO₂max is the point at which oxygen consumption plateaus despite anincrease in exercise intensity as shown on the chart in FIG. 11E.Alternatively, VO₂ max can be estimated using a number of physicaltests, including but not limited to, the Uth-Sorensen-Overgaard-Pedersenestimation, the Cooper test, the Multi-stage fitness test (or beeptest), a step test (e.g., Harvard Step Test, Queens College Step Test,Tecumseh Step Test), Storer Maximal Cycle Test, or the Rockport fitnesswalking test. VO₂ max can also be predicted using an exercise historyquestionnaire, age, height, weight, and gender (e.g., Jackson) or aperceived functional ability, physical activity rating scale, height,weight, and gender (e.g., George). In an alternative embodiment, VO₂ maxis estimated using data obtained from GPS devices, heart rate monitors,and fitness bands (e.g., UA Band, Garmin, Polar, Fitbit, TomTom, Soleus,Timex, Mio, Jawbone, Zephyr), mobile devices (e.g., iPhone, Android),and/or social networks and mobile applications (e.g., Record by UnderArmour, MapMyRun, MapMyRide, MapMyFitness, Endomondo, Strava,dailymile). VO₂ max can be estimated by using factors including but notlimited to maximum heart rate, resting heart rate, distance run in aspecific time, time to run a specific distance, gender, age, bodyweight, body mass, number of times a user exercises per week, and heartrate after exertion. In an alternative embodiment, VO₂ max is estimatedusing expected values, which are dependent on age, gender, and aself-reported rating, as shown in FIG. 11E. VO₂ max decreasesapproximately 0.54 mL/kg/min per year for women and approximately 0.35mL/kg/min per year in men.

Other types of manually input data include metabolic disorder, such asdiabetes. Since Type 1 diabetes is associated with reduced eccrine glandactivation and, thus, lower sweat rates, the present invention mayreveal user metabolic disorder. Further, automatically input dataincludes user skin temperature, outdoor or indoor temperature, time,date, humidity, and/or altitude. This data is also input manually inanother embodiment. Other automatically input data and/or manually inputdata includes exertion levels and/or body mass. The automatically inputdata may be generated in the remote transceiver device by integratedapplications, such as GPS or weather. Together, the data transmitted tothe microcontroller from the remote transceiver device representmodifying variables. Preferably, microcontroller software and/orsoftware on the computing device is operable to compensate forvariations across the automatically and/or manually input data.

The microcontroller converts the voltage data from the biological fluidinto a concentration or ratio value of the biomarker using at least oneprogrammed algorithm. For example, as shown in FIG. 11A, if thealgorithm was converting the amount of sodium ions detected at thesensor into a sodium concentration, the algorithm would apply around0.242 mM per mV of sodium. For potassium conversion, the ratio would bearound 0.195 mM per mV of potassium. The article Excretion of Sodium andPotassium in Human Sweat, by Schwartz, et al. in J. Clin. Invest. 1956January; 35(1); 114-120, which is incorporated by reference herein inits entirety, examines the concentrations of sodium and potassium inhuman sweat following administration of a cholinergic drug. In oneembodiment, sensor data are inputs into real time blood serum hydration,sodium concentration, and potassium concentration using absorption andextraction models that use sensor data as starting points. Thesecalculated values are the apparatus' output data. Types of output datainclude but are not limited to concentrations, such as molarity,osmolarity, and osmolality, and descriptive statistics, such asaverages, ratios, and trends, all of which may be categorized based on asub-range within a larger physiological range of the biomarker, as shownin FIG. 13A. The modifying variables transmitted from the remotetransceiver device may modify the algorithm, which may adjust the outputdata.

The output data is then transmitted from the apparatus to a remotecomputer device, such as by way of example and not limitation, asmartphone, a tablet computer, or wearable computer, preferably, throughwireless network communication by the transceiver antenna (which mayinclude a coil) of the apparatus. Using a larger antenna in the presentinvention provided for lower data loss and easier reads associated witha broad x-y placement tolerance. The wireless transmission is providedby any suitable wireless communication, wireless network communication,standards-based or non-standards-based, by way of example and notlimitation, radiofrequency, Bluetooth, Zigbee, Wi-Fi, near fieldcommunication, or other similar commercially utilized standards. At theremote transceiver device, the output data can be viewed and assessed bythe one or multiple users. The one or more users also may manipulate orfurther analyze the output data, such as by creating user defined graphsand tables. Preferably, the remote transceiver device is portable. Morepreferably, the device is a smartphone. Alternative devices include bulkreaders, such as food and or beverage dispensers with sensor and/ormobile app communication capabilities or athletic training gearincluding treadmills, spin bikes, ellipticals, stair climbers, andweight machines with integrated mobile communication capabilities. Morealternative devices include desktop or laptop computers and tablets.FIG. 16 diagrams the communication between the sensor apparatus and thesmartphone or reader, wherein power, commands, and/or data may becommunicated.

From the remote transceiver device, the user may transmit processed orunprocessed data to at least one remote computer server, preferably bywireless communication, such as through a user web service. The remotecomputer server, which may be a network or cloud, may store thetransmitted data in a library. The cloud preferably serves as a softwaredevelopment kit (SDK) for potential solution partners, a cloud baseduser app (with real time ingestion, calculation, and display), and acloud based user store with ubiquitous access.

The library includes functions, such as file storage, security,extensions, utilities, scheduling, messaging, persistence, cache, andlogging. FIG. 29 shows a cloud enterprise, wherein a cloud computingplatform receives data from application users and processes it forinternal and partner use. The software code that resides in acloud-based computer system of the present invention is designed,constructed, and configured to handle the unique data in unique ways. Itautomatically validates data (to determine if it is reasonable/useabledata) and triggers a series of workflows based on the type, date/timestamps, and scope of data to correlate and identify trends. It furtherincludes correlation and trending tags for subsequent useralerts/analysis. The code has an open framework built on web serviceconcepts to interact and integrate with other 3^(rd) party analytics.These web service calls are a series of open Application ProgrammingInterfaces (APIs) aggregated into a Software Development Kit (SDK) whichenables authorized 3^(rd) party developers to create and maintain 3^(rd)party user apps that leverage the cloud infrastructure to access/sharedata, correlations, trends, and other analytic results.

The two-way communication between the apparatus and the remotetransceiver device is significant for the fullness of systemfunctionality. As shown in FIG. 16 , the remote transceiver device maycommunicate with the apparatus to provide, by way of example and notlimitation, commands, electrode calibration, microcontroller softwareupdates, new or updated algorithms, and/or new or updated modifyingvariables for algorithms. Communication may be manually or audiblyactivated. The apparatus may communicate with the remote transceiverdevice to provide, by way of example and not limitation, output data,microcontroller health properties, error codes, electrode maintenance ormalfunction. At the transceiver device, the one or more users mayseparately or simultaneously view selected session tables, full historysession tables, sensor or multi-sensor chronology, and external sensorcorrelation. Further, selected biomarker or multi-biomarker historiesmay be viewed.

The system architecture is diagrammed in FIG. 20 . Here, the remotetransceiver device is characterized as a Mobile App, preferably on asmartphone, which is in communication with the sensor apparatus viawireless communication. Mobile App controls and commands are diagrammedin FIG. 21 . The Mobile App may network with the user web service, asshown in FIG. 22 , to access the cloud database and library, as shown inFIG. 23 . An example cloud platform operable with the present inventionis the EMITTI platform (ex: Amazon Web Services, Microsoft Azure, or anyother similar commercial or private cloud platform); cloud architectureis more specifically detailed in FIG. 28 . The web service allows theuser to access, analyze, and manipulate user output data that wastransmitted from the sensor apparatus. FIG. 24 shows an individual userweb front including features such as external sensor correlation,selected session summary, and session tables. FIG. 25 shows the user webfront for multiple users. The complete website builder platform is shownin FIG. 26 . The website may connect to the cloud computing enterprise,which is diagrammed in FIGS. 29 and 30 , or link to social media sites.

From the cloud computing system, data from multiple users may be stored,as diagrammed in FIG. 31 . Access to these data may be acquired byresearchers and epidemiologists to perform a variety of researchanalytics. The captured data will preferably be from the same sweatdetection model, providing greater reliability to the pool of data. Theability to collect these specific biomarker data from such a largepopulation of subjects creates an invaluable real-time, continuousepidemiological research system and method.

The preferred embodiment of the system includes an apparatus thatintimately adheres to mammalian skin, more specifically to human skin.The sweat from the skin is moved into the apparatus for detection ofsweat biomarkers and analytes. Where on the mammal the apparatus ispositioned is dependent upon, by way of example and not limitation, userpreference, sweat collection patterns, or sweat production amounts at agiven location.

The apparatus is operable to determine a measured amount oftransepidermal sweat and/or a measured amount of evaporative sweat andan estimated amount of transepidermal sweat and/or an estimated amountof evaporative sweat. In one embodiment, estimated amounts aredetermined based on body surface area, mass, gender, fitness level,weight, and/or age. In another embodiment, the apparatus is operable tocompare the estimated amount of transepidermal sweat and/or theestimated amount of evaporative sweat to the measured amount oftransepidermal sweat and/or the measured amount of evaporative sweat andprovide a status based on the comparison of the estimated amount oftransepidermal sweat and/or the estimate amount of evaporative sweat tothe measured amount of transepidermal sweat and/or the measured amountof evaporative sweat. In another embodiment, the apparatus uses atangible/quantifiable fitness level in combination with sweat biomarkerratios in order to calculate real-time sweat rates. In anotherembodiment, the apparatus uses gender factors in order to improve sweatflow rate accuracy. In another embodiment, the apparatus usesconsumption refresh models, exact custom formula to return to startcondition.

The apparatus is also operable to model losses and consumption of sweatin order to estimate blood serum characteristics at a time before use,at the start of use, in real-time, or at a time after use. In oneembodiment, the apparatus is operable to predict performance erosion andinjury probability based on the analysis of at least one biologicalfluid biomarker. In another embodiment, the apparatus is operable to usethe analysis of sweat to provide corrective action recommendations.

The basic fabrication process, as shown in FIG. 32 , includes thefollowing steps: substrate fabrication, circuit fabrication, pick andplace, reflow soldering, electrode fabrication, circuit boardprogramming, membrane fabrication, sealing and curing, and dressing.

Electrode fabrication includes the described novel metallization pasteand sequence for the reference probe; novel metallization applicationand sequence for active probes; novel line and space characteristics forsweat flow rate and small protein probes.

Membrane fabrication entails the described novel precision ionophoreapplication and cure processes.

Dressing entails the described novel laser cutting, bonding, andassembly steps for fabricating the microfluidic components and thedressing.

FIG. 33 is a schematic diagram of an embodiment of the inventionillustrating a cloud-based computer system, generally described as 800,having a network 810, a plurality of computing devices 820, 830, 840, aserver 850 and a database 870.

The server 850 is constructed, configured and coupled to enablecommunication over a network 810 with a computing devices 820, 830, 840.The server 850 includes a processing unit 851 with an operating system852. The operating system 852 enables the server 850 to communicatethrough network 810 with the remote, distributed user devices. Database870 may house an operating system 872, memory 874, and programs 876.

In one embodiment of the invention, the system 800 includes acloud-based network 810 for distributed communication via a wirelesscommunication antenna 812 and processing by a plurality of mobilecommunication computing devices 830. In another embodiment of theinvention, the system 800 is a virtualized computing system capable ofexecuting any or all aspects of software and/or application componentspresented herein on the computing devices 820, 830, 840. In certainaspects, the computer system 800 may be implemented using hardware or acombination of software and hardware, either in a dedicated computingdevice, or integrated into another entity, or distributed acrossmultiple entities or computing devices.

By way of example, and not limitation, the computing devices 820, 830,840 are intended to represent various forms of digital computers 820,840, 850 and mobile devices 830, such as a server, blade server,mainframe, mobile phone, a personal digital assistant (PDA), a smartphone, a desktop computer, a netbook computer, a tablet computer, aworkstation, a laptop, and other similar computing devices. Thecomponents shown here, their connections and relationships, and theirfunctions, are meant to be exemplary only, and are not meant to limitimplementations of the invention described and/or claimed in thisdocument

In one embodiment, the computing device 820 includes components such asa processor 860, a system memory 862 having a random access memory (RAM)864 and a read-only memory (ROM) 866, and a system bus 868 that couplesthe memory 862 to the processor 860. In another embodiment, thecomputing device 830 may additionally include components such as astorage device 890 for storing the operating system 892 and one or moreapplication programs 894, a network interface unit 896, and/or aninput/output controller 898. Each of the components may be coupled toeach other through at least one bus 868. The input/output controller 898may receive and process input from, or provide output to, a number ofother devices 899, including, but not limited to, alphanumeric inputdevices, mice, electronic styluses, display units, touch screens, signalgeneration devices (e.g., speakers) or printers.

By way of example, and not limitation, the processor 860 may be ageneral-purpose microprocessor (e.g., a central processing unit (CPU)),a graphics processing unit (GPU), a microcontroller, a Digital SignalProcessor (DSP), an Application Specific Integrated Circuit (ASIC), aField Programmable Gate Array (FPGA), a Programmable Logic Device (PLD),a controller, a state machine, gated or transistor logic, discretehardware components, or any other suitable entity or combinationsthereof that can perform calculations, process instructions forexecution, and/or other manipulations of information.

In another implementation, shown as 840 in FIG. 33 , multiple processors860 and/or multiple buses 868 may be used, as appropriate, along withmultiple memories 862 of multiple types (e.g., a combination of a DSPand a microprocessor, a plurality of microprocessors, one or moremicroprocessors in conjunction with a DSP core).

Also, multiple computing devices may be connected, with each deviceproviding portions of the necessary operations (e.g., a server bank, agroup of blade servers, or a multi-processor system). Alternatively,some steps or methods may be performed by circuitry that is specific toa given function.

According to various embodiments, the computer system 800 may operate ina networked environment using logical connections to local and/or remotecomputing devices 820, 830, 840, 850 through a network 810. A computingdevice 830 may connect to a network 810 through a network interface unit896 connected to the bus 868. Computing devices may communicatecommunication media through wired networks, direct-wired connections orwirelessly such as acoustic, RF or infrared through an antenna 897 incommunication with the network antenna 812 and the network interfaceunit 896, which may include digital signal processing circuitry whennecessary. The network interface unit 896 may provide for communicationsunder various modes or protocols.

In one or more exemplary aspects, the instructions may be implemented inhardware, software, firmware, or any combinations thereof. A computerreadable medium may provide volatile or non-volatile storage for one ormore sets of instructions, such as operating systems, data structures,program modules, applications or other data embodying any one or more ofthe methodologies or functions described herein. The computer readablemedium may include the memory 862, the processor 860, and/or the storagemedia 890 and may be a single medium or multiple media (e.g., acentralized or distributed computer system) that store the one or moresets of instructions 900. Non-transitory computer readable mediaincludes all computer readable media, with the sole exception being atransitory, propagating signal per se. The instructions 900 may furtherbe transmitted or received over the network 810 via the networkinterface unit 896 as communication media, which may include a modulateddata signal such as a carrier wave or other transport mechanism andincludes any delivery media. The term “modulated data signal” means asignal that has one or more of its characteristics changed or set in amanner as to encode information in the signal.

Storage devices 890 and memory 862 include, but are not limited to,volatile and non-volatile media such as cache, RAM, ROM, EPROM, EEPROM,FLASH memory or other solid state memory technology, disks or discs(e.g., digital versatile discs (DVD), HD-DVD, BLU-RAY, compact disc(CD), CD-ROM, floppy disk) or other optical storage, magnetic cassettes,magnetic tape, magnetic disk storage or other magnetic storage devices,or any other medium that can be used to store the computer readableinstructions and which can be accessed by the computer system 800.

It is also contemplated that the computer system 800 may not include allof the components shown in FIG. 33 , may include other components thatare not explicitly shown in FIG. 33 , or may utilize an architecturecompletely different than that shown in FIG. 33 . The variousillustrative logical blocks, modules, elements, circuits, and algorithmsdescribed in connection with the embodiments disclosed herein may beimplemented as electronic hardware, computer software, or combinationsof both. To clearly illustrate this interchangeability of hardware andsoftware, various illustrative components, blocks, modules, circuits,and steps have been described above generally in terms of theirfunctionality. Whether such functionality is implemented as hardware orsoftware depends upon the particular application and design constraintsimposed on the overall system. Skilled artisans may implement thedescribed functionality in varying ways for each particular application(e.g., arranged in a different order or partitioned in a different way),but such implementation decisions should not be interpreted as causing adeparture from the scope of the present invention.

By way of definition and description supporting the claimed subjectmatter, preferably, the present invention includes communicationmethodologies for transmitting data, data packets, messages or messagingvia a communication layer. Wireless communications over a network arepreferred. Correspondingly, and consistent with the communicationmethodologies for transmitting data or messaging according to thepresent invention, as used throughout this specification, figures andclaims, wireless communication is provided by any reasonable protocol orapproach, by way of example and not limitation, Bluetooth, Wi-Fi,cellular, ZigBee, near field communication, or other similarcommercially utilized standards; the term “ZigBee” refers to anywireless communication protocol adopted by the Institute of Electronics& Electrical Engineers (IEEE) according to standard 802.15.4 or anysuccessor standard(s), the term “Wi-Fi” refers to any communicationprotocol adopted by the IEEE under standard 802.11 or any successorstandard(s), the term “WiMax” refers to any communication protocoladopted by the IEEE under standard 802.16 or any successor standard(s),and the term “Bluetooth” refers to any short-range communicationprotocol implementing IEEE standard 802.15.1 or any successorstandard(s). Additionally or alternatively to WiMax, othercommunications protocols may be used, including but not limited to a“1G” wireless protocol such as analog wireless transmission, firstgeneration standards based (IEEE, ITU or other recognized worldcommunications standard), a “2G” standards based protocol such as “EDGEor CDMA 2000 also known as 1×RTT”, a 3G based standard such as “HighSpeed Packet Access (HSPA) or Evolution for Data Only (EVDO), anyaccepted 4G standard such as “IEEE, ITU standards that include WiMax,Long Term Evolution “LTE” and its derivative standards, any Ethernetsolution wireless or wired, or any proprietary wireless or power linecarrier standards that communicate to a client device or anycontrollable device that sends and receives an IP based message. Theterm “High Speed Packet Data Access (HSPA)” refers to any communicationprotocol adopted by the International Telecommunication Union (ITU) oranother mobile telecommunications standards body referring to theevolution of the Global System for Mobile Communications (GSM) standardbeyond its third generation Universal Mobile Telecommunications System(UMTS) protocols. The term “Long Term Evolution (LTE)” refers to anycommunication protocol adopted by the ITU or another mobiletelecommunications standards body referring to the evolution ofGSM-based networks to voice, video and data standards anticipated to bereplacement protocols for HSPA. The term “Code Division Multiple Access(CDMA) Evolution Date-Optimized (EVDO) Revision A (CDMA EVDO Rev. A)”refers to the communication protocol adopted by the ITU under standardnumber TIA-856 Rev. A.

It will be appreciated that embodiments of the invention describedherein may be comprised of one or more conventional processors andunique stored program instructions that control the one or moreprocessors to implement, in conjunction with certain non-processorcircuits, some, most, or all of the functions for the systems andmethods as described herein. The non-processor circuits may include, butare not limited to, radio receivers, radio transmitters, antennas,modems, signal drivers, clock circuits, power source circuits, relays,current sensors, and user input devices. As such, these functions may beinterpreted as steps of a method to distribute information and controlsignals between devices. Alternatively, some or all functions could beimplemented by a state machine that has no stored program instructions,or in one or more application specific integrated circuits (ASICs), inwhich each function or some combinations of functions are implemented ascustom logic. Of course, a combination of the two approaches could beused. Thus, methods and means for these functions have been describedherein. Further, it is expected that one of ordinary skill in the art,notwithstanding possibly significant effort and many design choicesmotivated by, for example, available time, current technology, andeconomic considerations, when guided by the concepts and principlesdisclosed herein, will be readily capable of generating such softwareinstructions, programs and integrated circuits (ICs), and appropriatelyarranging and functionally integrating such non-processor circuits,without undue experimentation.

Certain modifications and improvements will occur to those skilled inthe art upon a reading of the foregoing description, by way of example,a device having at least one microprocessor for storing data may beoperable in the device before data transmission. Another exampleincludes other advanced sensors, as well as being incorporated intosmart fabrics and protective wear. Advanced sensors include advancedsweat biomarkers, pulse rate, breath rate, micro EKG, micro O2, picturelog, voice log, voice translate, tissue safe X-ray, blood pressure, andcombinations thereof. Smart fabrics incorporate the present inventionand include active heat/cooling, kinetic energy generation,electromagnetic energy harvesting, wearable energy storage, wearabledata storage, wearable processing, wearable communications, elastomericactuators, and combinations thereof. More generally, the apparatus maybe part of apparel and material for lower body clothing and upper bodyclothing. The present invention is also incorporated into enhancedprotective wear such as enhanced helmets, gloves and footwear.Specifically, sensors, conductors and/or ionophores are utilized onmoisture management fabrics, such as by way of example and notlimitation, Under Armour fabrics. Microfluidic moisture transport isalso utilized in fabric and other material which directly contacts humanskin when worn. Enhanced helmets include those with MM, 3D audio, visualenhancement, mixed reality, breath sensors, aerosol nutrition andcombinations thereof. Enhanced gloves include touch communications,elastomeric grip, gesture control and combinations thereof. Enhancedfootwear includes power generation boots, 3D tracking, tactile alerts,communication transceivers, and combinations thereof.

The above mentioned examples are provided to serve the purpose ofclarifying the aspects of the invention and it will be apparent to oneskilled in the art that they do not serve to limit the scope of theinvention. All modifications and improvements have been deleted hereinfor the sake of conciseness and readability but are properly within thescope of the present invention.

What is claimed is:
 1. A method of fabrication for a biological fluidsensor for analyzing at least one analyte in human biological fluid,said biological fluid sensor comprising electronic components,electrochemical components, and chemical components, the methodcomprising: fabricating at least one substrate; fabricating at least onecircuit on the at least one substrate; placing the electronic componentsonto the at least one circuit, wherein the electronic components includeat least one microcontroller and at least one transceiver; fabricatingat least two electrodes, wherein the at least two electrodes arecomprised of at least one active electrode and at least one referenceelectrode; applying an ionophore coating on the at least one activeelectrode; sealing the at least one active electrode to create anelectronic layer; and integrating the electronic layer with a top coverlayer, a microfluidic management layer, and a double-sided adhesivelayer; wherein the step of fabricating the at least two electrodesfurther comprises: fabricating a metallization paste; constructing theat least one reference electrode by applying the metallization paste ontop of at least one first trace metal of the at least one circuit andannealing; and constructing the at least one active electrode byapplying the metallization paste on top of at least one second tracemetal of the at least one circuit and annealing; forming a conductivetrace in a first ring around the at least one active electrode, whereinthe conductive trace in the first ring around the at least one activeelectrode does not contact the at least one active electrode; forming asecond ring around the at least one active electrode, wherein the secondring around the at least one active electrode is inside the first ringaround the at least one active electrode, and wherein the first ringaround the at least one active electrode does not contact the secondring around the at least one active electrode; and applying theionophore coating on the at least one active electrode such that theionophore coating completely covers the at least one active electrodeand is contained within the second ring; wherein the step of integratingthe electronic layer with the top cover layer, the microfluidicmanagement layer, and the double-sided adhesive layer further comprises:placing the electronic layer on top of an adhesive side of the top coverlayer; surrounding the at least two electrodes with the microfluidicmanagement layer; placing a double-sided adhesive layer on top of theelectronic layer such that the double-sided adhesive layer covers theelectronic layer; and pressing the top cover layer, the microfluidicmanagement layer, the electronic layer, and the double-sided adhesivelayer.
 2. The method of claim 1, wherein the biological fluid is sweat.3. The method of claim 1, wherein the metallization paste is made of atleast one stable metal selected from the group consisting of silver,gold, platinum, and palladium.
 4. The method of claim 1, wherein the atleast one reference electrode and the at least one active electrodeinclude silver, zinc, copper, gold, platinum, rhodium, carbon, or acombination thereof.
 5. The method of claim 1, wherein applying theionophore coating on the at least one active electrode is performed viaan automated dispenser.
 6. The method of claim 1, further includingcuring the ionophore coating using heat or light.
 7. The method of claim1, further including reflow soldering of the at least one circuit. 8.The method of claim 1, wherein the ionophore coating is fabricated bydissolving an ion selective material in a polyvinylchloride (PVC)solution.
 9. The method of claim 1, wherein the ionophore coating iscast in a shape and placed on a carrier or a film to facilitateapplication on the at least one electrode.
 10. The method of claim 9,wherein the carrier or the film becomes an integral part of thebiological fluid sensor.
 11. The method of claim 1, wherein theionophore coating is applied to the at least one active electrode usingan adhesive or a bonding film.
 12. The method of claim 1, wherein theionophore coating has an embedded glass or plastic fiber.
 13. The methodof claim 1, wherein the microfluidic layer is comprised of wickingpaper, fabric, or crystal fibers.
 14. The method of claim 1, wherein theat least one analyte is selected from the group including sodium,potassium, chloride, oxygen, glucose, calcium, ammonium, copper,magnesium, iron, zinc, lactate, creatinine, uric acid, urea, ethanol,amino acids, hormones, steroids, proteins, catecholamines, andinterleukins.
 15. The method of claim 1, wherein the double-sidedadhesive layer further contains laser cut macrofluidic pores.
 16. Themethod of claim 1, wherein the step of integrating the electronic layerwith the top cover layer, the microfluidic management layer, and thedouble-sided adhesive layer further includes: mechanically separating anindividual unit; and automatically packaging the individual unit into anindividualized wrapper to create an individually wrapped unit.